Objective To investigate the protective mechanism of ischemic preconditioning on ischemic/reperfusion injury after pancreas transplantation in rats.Methods The model of diabetic SD rat was established. Twenty-four diabetic SD rats were randomly assigned to ischemic/reperfusion group (I/R group, n=6) and ischemic preconditioning group (IPC group, n=18). The rats in group IPC were averagely assigned to 3 sub-groups: group IPC_ 1 (5 min ischemic and 5 min reperfusion), IPC_ 2 (5 min ischemic and 5 min reperfusion twice) and IPC_ 3 (5 min ischemic and 5 min reperfusion thrice). Six normal SD rats whose abdomen was opened only served as control group, and they did not receive pancreas transplantation. I/R group and IPC group received pancreas transplantation. The superoxide dismutase (SOD) and myeloperoxidase (MPO) activity of grafts were monitored 2 h after reperfusion, the apoptotic cells in grafts were observed by TUNEL method, and the expression of Bcl-2 and Bax gene of the grafts was detected by Western blot.Results As compared with I/R group, the SIOD activity and the expression of Bcl-2 gene of grafts were significantly increased, while MPO activity, apoptotic index and the expression of the Bax gene in the grafts were markedly reduced in IPC group (P

Objective To study the effects of octreotide on the proliferation and apoptosis of hepatocellular carcinoma cells, and provide an experimental basis for clinical application. Methods The proliferation of hepatocellular carcinoma cells (HepG2) was assessed by MTT and growth curve respectively, the contents of AFP in the culture supernatant were determined by eletrochemiluminescence immunoassay, and apoptosis was detected by fluorescent staining, transmission electron microscopy and flow cytometry. Results The proliferation of HepG2 was inhibited significantly by octreotide with a dosage dependant manner(range from 0.005 to 80 ?g/ml, P

Objective To probe into the optimal concentration of Wnt-11 to induce the differentiation of bone marrow mesenchymal stem cells (BMMSCs)into cardiomyocyte-like cells in vitro.Methods BMMSCs were isolated from the bone marrow of SD rats using whole bone marrow culture method.After cultured for 48 h, BMMSCs of the second generation were utilized for directed induction.Based on the final concentration of Wnt-11 , BMMSCs were divided up into Group A (100 ng/mL),Group B (200 ng/mL),Group C (400 ng/mL)and Group D (blank control).After 72-hour induction,the cells were cultured in complete medium for 4 weeks while cells in Group D were cultured only in the complete medium.The morphological changes were observed under the phase contrast microscope.Surface antigen expression of BMMSCs was identified by flow cytometry.When cells were cultured for 4 weeks,the expressions of Desmin,Connexin43 and cTnI were detected by immunocytochemistry. Meanwhile, the ultrastructural changes were observed using transmission electron microscope. The mRNA expressions of cardiac transcription factors GATA-4,Nkx2.5 andα-MHC in BMMSCs were detected by RT-qPCR at 1,2 and 4 weeks after induction.Results Primary BMMSCs formed cell colonies at 2 weeks;the cells were mainly fusiform or star-shape,and a few irregularly-shaped ones were also visible.The passaged cells were larger than those of primary culture.After induction,the cells exhibited long shuttle-shape and were aligned in parallel. Flow cytometery displayed that the positive rate of the surface antigens of BMMSCs CD29,CD45,and CD90 was 97.9%,0.4% and 99.5%,respectively.When BMMSCs-induced via Wnt-11 were cultured for 4 weeks,Desmin, cTnI and Connexin43 were all positively expressed in induction groups.Whereas in the blank control group they were slightly positive or negative;the positive rate in Group B was the highest (P<0 .05 ).Transmission electron microscopy exhibited that organelles such as rough endoplasmic reticulum,mitochondria,as well as some ribosomes were visible in the cytoplasm of these cells in each induction group.In addition,myofilaments were arranged in parallel in the cytoplasm.The cells in induction groups could express GATA-4 and Nkx2 .5 in the first week,and then the expression of them decreased in the second week,but then increased in the fourth week;gene expression in induction Group B was significantly higher than in the other two induction groups (P<0 .05 ).The expression of GATA-4 and Nkx2 .5 in Group D was 1 ,α-MHC was not expressed in the four groups during the culture period. Conclusion Wnt-11 can induce the differentiation of BMMSCs into cardiomyocyte-like cells in vitro,and the optimal concentration of Wnt-11 is 200 ng/mL.

BACKGROUND:Ischemia/reperfusion (IR)injury during the pancreas transplantation can cause numerous postoperative complications, among which,secondary pancreatitis can cause small intestinal mucosal injury and result in severe Consepuence.OBJECTIVE:To observe the protective effect of ischemic preconditioning (IPC) on small intestinal mucosal barrier after pancreas transplantation in rats.DESIGN:Randomized controlled animal trial.SETTING:Department of Gastrointestinal Surgery,Xijing Hospital,Fourth Military Medical University of Chinese PLA.MATERIALS:This trial was done in the Laboratory of Gastrointestinal Surgery,Xijing Hospital,Fourth Military Medical University of Chinese PLA between September 2001 and April 2004.Eighty-three male SD rats were involved in this trial.METHODS: Forty-seven rats were randomly chosen to prepare diabetic rat models by penile-intravenous injection of 65 mg/kg streptozotocin.Thirty-six successful model rats were randomized into 3 groups,with 12 in each group:IR group,donor IPC(DIPC)group and recipient with two hindlims IPC(RIPC)group.Twelve of the remaining 36 normal rats served as control group,and the other 24 rats were used as donors.Laparotomy was conducted only in control group,and pancreas transplantation was conducted in the other 3 groups In DIPC group,the splenic vessels of donors were blocked for 5 minutes and reperfused for 5 minutes twice before obtaining pancreas from donor;In the RIPC group, blood flow of two hindlimbs of recipients was blocked for 5 minutes and reperfused for 5 minutes before reperfusing the pancreas of donor,and this procedure was repeated 3 times.IR group was untouched.MAIN OUTCOME MEASURES:① On the 5th day after operation,6 rats were randomly chosen from each group to detect small intestinal permeability[expressed with plasm fluorescent-isothiocyanate-dextran(FITC-dextran)concentration]and absorption function(expressed with plasm xylose concentration).② On the 5th day after operation.blood was taken from the left 6 rats in each group to detect serum tumor necrosis factor-α(TNF-α) and nitric oxide(NO)level as well as superoxide dismutase(SOD)and amylase activity.Ileal mucosal tissue was taken to detect wet weight of small intestinal mucosa,the height and width of microvilli,malonaldehyde(MDA)level and myeloperoxidase(MPO)activity.At the same time,mesenteric lymph node,liver and splenic tissue were taken to perform bacterial culture.Bacterial translocation was observed.RESULTS:After supplement,72 rats were involved in the result analysis.①Plasm FITC-dextran concentration of IR group were higher than that in control group,DIPC group and RIPC group,respectively(P＜0.01).②Plasm xylose concentration in the IR group was lower than that in the control group,DIPC group and RIPC group,respectively(P＜0.01).③Bacterial translocation rate in the IR group was higher than that in the control group,DIPC group and RIPC group,respectively(P＜0.01).④Small intestinal mucosal injury degree in the IR group was lower than that in the other 3 groups(P＜0.01).⑤Small intestinal MPO activity and MDA level in IR group were significantly higher than those in the other 3 groups(P＜0.01). Serum SOD activity and NO level were lower but amylase activity and TNF-α 1evel were higher in the IR group as compared with the other 3 groups(P＜0.01).CONCLUSION:IPC of two hindlimbs in both donor and recipient can protect small intestinal mucosal barrier and reduce bacterial translocation rate after pancreas transplantation in rats.

BACKGROUND: Pancreas transplantation alone (PTA) is an effective therapy for diabetic patients who do not occur chronic complications. It's important to establish the stable PTA animal models to investigate immunologic tolerance or ischemic/reperfusion injury.OBJECTIVE: To establish the model of pancreas transplantation alone (PTA) with enteric drainage in rat.DESIGN: Grouping and controlled animal experiment SETTING: Department of Gastrointestinal Surgery, Xijing Hospital,Fourth Military Medical University of Chinese PLA MATERIALS: Totally 90 SD male rats, with the body mass of 250-320 g,were chosen in this study. 58 rats were induced by intravenous administration of streptozotocin (STZ) at a dose of 65 mg/kg via penile vein and the rats whose fasting plasma glucose exceeded 19.4 mmol/L for more than 2weeks were selected, 22 rats was successful. Rats were randomly assigned to 2 groups: control group (10 healthy rats) and group PTA consisted of 22diabetic rats, which received PTA from 22 normal donors.METHODS: This experiment was conducted at the laboratory of Department of Gastrointestinal Surgery, Xijing Hospital, Fourth Military Medical University of Chinese PLA from January 1999 to July 2004. The blood vessels reconstruction of PTA were performed using end-to-side anastomosis between the donors' abdominal aorta segment (abdominal artery and splenic artery) and recipients' abdominal aorta, and end-to-end anastomosis between the donors' portal vein segment (splenic vein) and recipients'left renal vein (use a cuff). Pancreas exocrine drainage was made by pancreas intestine anastomosis (Roux-Y).MAIN OUTCOME MEASURES: Body mass, food intake, water intake and fasting blood glucose were monitored 2 days before operation and 1,3,7,14 and 30 days after operation, and the failure causes were analyzed.RESULTS: 22 rats in the model group and 10 rats in the normal control the vein of the rats , very severe diabetic symptoms appeared in 22 rats:Body mass, food intake, fasting blood glucose was increased than that of cipients operation was (32.2±12.7) minutes and (63.4±15.9) minutes respectively. And the mean time of warm and cold ischemic time was 0minute and (48.6±18.3) minutes, respectively. 11 of the 22 cases (50%)died or lost their function of the endocrine within 1 month in Group PTA.The main complications were secondary pancreatitis and pancreas leakage after transplantation (7 cases, 31.8%). All successful recipients' blood glucose lowed on the 1st and recovered to be normal on the 3nd after transplantation (P ＜ 0.01), and their food intake, water intake and urine volume decreased and became stable 14 days later.CONCLUSION: This method can be used to establish relative stable animal model. Successful PTAs may improve the pancreatic endocrine function of the diabetic rats.

Objective To explore the effect of recombinant pEGFP-N3-APC vectors carrying various APC functional domains on the expression of β-catenin in human colorectal cancer cells HT-29.Methods The recombinant plasmids were transfected into HT-29 cells mediated by lipofectamine~(TM) 2000, and detected by green fluorescence and RT-PCR. Western blot was applied to detect β-catenin expression level in HT-29 cells after transfection, and gray scales of electrophoresis strips were analyzed by SPSS 13.0.Results Green fluorescence and RT-PCR made clear that all 5 recombinant plasmids were successfully expressed in HT-29 cells. Western blot showed that β-catenin expression level in HT-29 cells was not affected after being transfected with pEGFP-N3-APC1, pEGFP-N3-APC2 and pEGFP-N3-APC3, and was distinctly affected after being transfected with pEGFP-N3-APC4 and pEGFP-N3-APC5, especially the later one. Conclusion The selected APC5 gene fragment with 15-amino acid repeats and SAMP repeats, which is relatively short, can degrade β-catenin level in HT-29 cells and may be applied in the gene therapy.

Objective Tracking of the vaginal microflora recovery and the expression of immune factors from untreated and treated patients with bacterial vaginosis (BV) by using different administration regimens and studying the relationship of treatment results and regimen selections. Methods 25 healthy females were selected as a control group and 100 BV patients were randomly divided into 4 groups (n=25/group). Group A: Intravaginal administration of metronidazole (× 7 d), Group B:Continuous intravaginal administration of metronidazole (× 7 d) and then live Lactobacillus Capsule (× 7 d) , Group C: Intravaginal administration of nifuratel (× 7 d), Group D: Continuous intravaginal administration of nifuratel (× 7 d) and then live Lactobacillus Capsule (×7 d). The microecological assessment system and EILSA were used to compare the clinical efficacy, vaginal microflora recovery and the changes in IL-8, TLR2 and TNF-αof the vaginal lavage fluid in healthy women or patients with bacterial vaginosis before and after treatments by four treatment strategies. Results ① The vaginal microflora imbalance, flora disturbance, pH value increased were presented in BV group compared with the control group.②Compared to the median of IL-8, TLR2 and TNF-α in vaginal lavage fluids of control group, there was no significant difference in IL-8 level but both TLR2 and TNF-αwere significantly increased (P<0.05) in BV group. The immune factors had no significantly difference in all BV groups.③The therapeutic effect in each BV groups was compared after stopping treatment for 7 days. The cure rate and the vaginal microflora recovery rate were significant higher in group B and D than group A and C (P<0.05). ④ After treatment there was no significant change in IL-8 level but there was an obviouslydecrease in TLR2 and TNF-α(P<0.05). The decreased levels are more significant in groups B and D than groups A and C (P<0.05). Conclusion By combining with the microecological assessment system to evaluate the therapeutic effect of BV, our research suggests that the sequence schemes of nifuratel plus live Lactobacillus Capsule is more effective in therapy effect, restoring normal vaginal micro-ecological environment and vaginal local immunity than metronidazole used alone.

Objective To optimize the matrix formula of propranolol hydrochloride gels. Methods On the basis of single factor experiment, central composite design-response surface method was used to optimize the formula. Addition levels of glycerol (A), PEG-400 (B) and HPMC (C) were evaluated as the independent variables.Eight-hour accumulative penetration amount per unit area measured by HPLC was used as the index. Quadratic polynomial was used to estimate the relationship between the index and the independent variables, and to delineate response surface and overlay contour plots in order to select the optimal formulations.Finally, predicted responses were verified. Results The optimized formula consisted of 18.53% glycerol, 8.54% PEG-400 and 2.35% HPMC.The quadratic polynomial regression model of 8-h accumulative penetration amount per unit area:R1=-7 415. 69+306. 10A+167. 47B+4 820. 59C-8. 26A2-9. 81B2-1 025. 75C2 , and the cumulative transmittance was 49.6%. Conclusion A credible model is established by using central composite design-response surface method and the formula of propranolol hydrochloride gels is optimized and the gel is stable and controllable.

Objective To explore the effect on ultrasound localization method with radial artery cannulation in infants.Methods Sixty infants (1 month-1year old)undergoing elective surgical proce-dures required artery pressure,were randomly divided into two groups:ultrasound localization group (group B):the infants were used of portable B ultrasound probe in the wrist with two dimensional ul-trasound image to determine the body position of the radial artery;control group (group C):tradi-tional palpation determine the radial artery puncture position.One puncture success rate,total punc-ture success rate,the period of puncture time and incidence of complications were recorded;MAP, the radial artery diameter,the vertical distance from the ultrasound probe to the skin and the length from the puncture spot to the radial artery were also measured.Results The success rate of first puncture in group B was higher than that of group C (P <0.05),the times of puncture and procedure time were significantly less than those of group C (P <0.05),the incidence of hematoma was lower than that in group C (P <0.05).Conclusion The application of ultrasound-guided method for radial artery cannulation is safe and reliable with accurate localization,highly successful rate of puncture at first time,lower complication and shorter time for placement.

BACKGROUND:Biological dressing A (porcine Xenoderm) and Physiotule Ag (Coloplast) show good effects on the absorption of exudates, adhesion and anti-bacteria in treatment of degree Ⅱ facial burns. OBJECTIVE:To observe the effectiveness of biological dressing Aversus Physiotule Ag after early debridement of degree Ⅱ facial burns. METHODS:A total of 15 patients with superficial degree Ⅱ facial burn and 10 patients with deep degree Ⅱ facial burns were included. Symmetric facial area of the same patient with the same depth of burn was divided into two parts of same size for treatments. One side was randomly selected as the experimental side, and treated with biological dressing A. The other was selected as the control side and treated with Physiotule Ag. We compared wound healing time, infection, times of changing dressings, skin after healing, drug change comfort and dressing comfort between the two sides. RESULTS AND CONCLUSION:In patients with superficial grade Ⅱ facial burn, times of changing dressings and drug change comfort were better in the experimental side than in the control side (P < 0.05), but the dressing comfort was better in the control side than the experimental side (P< 0.05). No significant difference in wound healing time, infection and skin after healing was detected between the two sides. In patients with deep degree Ⅱ facial burns, wound healing time, times of changing dressings, skin after healing and drug change comfort were better in the experimental side than in the control side (P < 0.05), but the dressing comfort was better in the control side than the experimental side (P< 0.05). No significant difference in infection was detectable between the two sides. Above findings suggested that the therapeutic effects of biological dressing A and Physiotule Ag were similar in treatment of degree Ⅱ facial burns. Biological dressing A in repair of deep degree Ⅱ facial burns promotes the wound healing andelevates the quality of healing.